Objective: To investigate the expression of miR-33b in hepatocellular carcinoma (HCC) and to explore regulatory mechanism of miR-33b for cell proliferation of HCC.
Methods: HCC tissues and adjacent non-tumor tissues were collected for this study (n=32 for each). Real-time PCR and Western blot were conducted to examine the mRNA and protein expression, respectively. MTT assay was used to detect the cell proliferation. Luciferase reporter gene assay was performed to verify the target relationship between miR-33b and Sal-like 4 (SALL4).
Results: MiR-33b was significantly downregulated in HCC tissues compared with adjacent non-tumor tissues. Overexpression of miR-33b decreased the proliferation of HCC LH86 cells. SALL4 was identified as a target gene of miR-33b, and its protein expression was negatively regulated by miR-33b. Overexpression of SALL4 reversed the suppressive effect of miR-33b on LH86 cell proliferation. SALL4 was significantly upregulated in HCC tissues compared with adjacent non-tumor tissues.
Conclusion: The miR-33b suppresses HCC cell proliferation through down-regulation of SALL4.
目的：探索微小RNA-33b(micro RNA-33b，miR-33b)在肝细胞癌(hepatocellular carcinoma，HCC)中的表达及调控HCC细胞增殖的相关分子机制。方法：收集配对的HCC及癌旁组织32对，分别采用实时荧光定量 PCR和Western印迹法检测人类婆罗双树样基因4(Sal-like 4，SALL4)RNA和蛋白表达， MTT实验检测HCC细胞增殖，萤光素酶报告基因检测验证miR-33b与SALL4基因的靶点关系。结果：MiR-33b在HCC组织中的表达显著低于癌旁组织(P<0.001)。过表达miR-33b能显著降低HCC LH86细胞的增殖。SALL4是miR-33b的靶基因，其蛋白表达被miR-33b负调控。过表达SALL4能逆转miR-33b对LH86细胞增殖的抑制作用。SALL4在HCC组织中的表达显著高于癌旁组织(P<0.001)。结论：MiR-33b对HCC细胞增殖的抑制作用是通过负调控SALL4的表达而实现。.